Fig. 2. HPLC chromatograms of A. agallocha MeOH extract (left) and Hes1 binding natural products after screening (right). For screening, the amount of proteins on beads were controlled; GST-Hes1 beads (GST-Hes1: ca. 3.6 nmol), GST-beads (GST: ca. 3.8 nmol). The mixture of beads (bed volume 100 μl) and extract (125 μg in EtOH, 25 μl) was incubated at 4 °C for 2 h. After washing, the binding natural products were dissociated from proteins by addition of 70% EtOH and heating (100 °C, 3 min).
